To assess the cell cycle profile, the cells were treated with butein and were subsequently fixed in 95% ethanol with 0.5 % Tween-20 at −20°C overnight. The fixed cells were stained with 50 µg/ml propidium iodide (Santa Cruz Biotechnology, Inc.). For apoptotic cell death, the cells were treated with butein and subsequently stained with annexin V and 7-aminoactinomycin D (7-AAD; Santa Cruz Biotechnology, Inc.). The cells were analyzed using a FACSCalibur (BD Biosciences, San Jose, CA, USA) with CellQuest Pro, version 5.2 software.
7 aminoactinomycin d 7 aad
Manufactured by Santa Cruz Biotechnology
7-aminoactinomycin D (7-AAD) is a fluorescent dye used in flow cytometry applications. It binds to DNA and can be used to stain and detect dead or dying cells.
Automatically generated - may contain errors
2 protocols using 7 aminoactinomycin d 7 aad
1
Cell Cycle and Apoptosis Analysis via Flow Cytometry
2
Cell Viability Assay via Flow Cytometry
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Cell were trypsinized,
centrifuged at 500g for 5 min, resuspended in PBS,
and stained with 7-aminoactinomycin D (7AAD, Santa Cruz Biotechnology),
a DNA intercalating dye excluded from viable cells due to membrane
impermeability. The 7AAD was excited at 546 nm and analyzed at 656
nm. Only apoptotic and necrotic cells are stained by these dyes. After
incubation for 50 min in the dark at 37 °C, the cells were analyzed
by flow cytometry (CytoFlex, Beckman Coulter). The percentages of
cells stained with or without dyes were analyzed using the CytExpert
Software (Beckman Coulter).
centrifuged at 500g for 5 min, resuspended in PBS,
and stained with 7-aminoactinomycin D (7AAD, Santa Cruz Biotechnology),
a DNA intercalating dye excluded from viable cells due to membrane
impermeability. The 7AAD was excited at 546 nm and analyzed at 656
nm. Only apoptotic and necrotic cells are stained by these dyes. After
incubation for 50 min in the dark at 37 °C, the cells were analyzed
by flow cytometry (CytoFlex, Beckman Coulter). The percentages of
cells stained with or without dyes were analyzed using the CytExpert
Software (Beckman Coulter).
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